Delineation of erythropoiesis in normal and malignant bone marrow using monoclonal antibody AS-E1 directed against transferrin receptors (CD71).
We have delineated the erythropoietic compartment in normal and malignant bone marrow (BM) by using the monoclonal antibody (mAb) AS-E1 directed against the transferrin receptor by flow cytometric (FCM) analysis. In normal BM we found a bimodal expression in antigen density with a minor subset (approximately 3%) expressing AS-E1high and a larger subset (approximately 15%) expressing AS-E1low. By fluorescence activated cell sorting, morphological examination of smears stained by immunocytochemistry and by BFU-E assays the AS-E1high fraction was shown to contain cells of erythroid origin (proerythroblasts, basophilic erythroblasts and polychromatic erythroblasts), whereas the AS-E1low fraction consisted mainly of promyelocytes and myelocytes. In patients with malignant hematological disorders we found a more pronounced heterogeneity in the density and the degree of AS-E1low expression compared to normal BM, and to further characterize the AS-E1low cells in patients and to exclude that this broad reactivity interfered with the identification of the AS-E1high cells, we employed triple-color FCM assays with mAbs directed against the myeloid surface markers CD13 and CD66 in addition to AS-E1. In all patients we found that 80-90% of the AS-E1low cells co-expressed CD13 and/or CD66 and thus were of myeloid origin. Finally, we evaluated 2 methods for determination of the AS-E1high subset and found an assay involving forward light scatter and logAS-E1 density to be sufficient. We conclude that AS-E1high is a valid FCM marker for the normal erythropoiesis.
Olesen
G
G
Department of Haematology and Medicine, Aarhus University Hospital, Denmark.
Carlsen
I
I
Skovbo
A
A
Hokland
M
M
Hokland
P
P
D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology*; D014408:Biomarkers, Tumor; D001853:Bone Marrow / Q000502:physiology* / Q000503:physiopathology; D019046:Bone Marrow Neoplasms / Q000503:physiopathology*; D002469:Cell Separation; D004920:Erythropoiesis; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D019891:Leukopoiesis; D051379:Mice; D008807:Mice, Inbred BALB C; D011990:Receptors, Transferrin / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D014408:Biomarkers, Tumor; C068126:CD71 antigen; D011990:Receptors, Transferrin